ObjectiveTo investigate the effect of icariin （ICA） on steroid-induced ferroptosis in bone microvascular endothelial cells （BMECs）. MethodsRat BMECs were selected and treated with 500 mg·L^-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group， hormone group （500 mg·L^-1 hydrocortisone）， ICA group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA）， and ferroptosis agonist group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA + 2.7 mg·L^-1 erastin）. Cell viability was detected by CCK-8. The levels of ferrous ion， glutathione （GSH）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and reactive oxygen species （ROS） were detected by related kit species. The ferroptosis-related proteins， such as glutathione peroxidase 4（GPX4）， ferritin light chain （FTL）， and transferrin receptor protein1 （sTfR） were detected by Western blot， as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B （LC3B）， Beclin1， B-cell lymphoma-2 （Bcl-2）， and Caspase-3. Results500 mg·L^-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs， and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity， compared with those in the blank group， the cell vitality， GSH in the hormone group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions were significantly increased （P<0.01）. Compared with those in the hormone group， the cell viability， GSH were significantly increased， and the levels of ROS， SOD， MDA， and ferrous ions were decreased in the ICA group （P<0.01）. Compared with those in the ICA group， the cell vitality， GSH in the ferroptosis agonist group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions increased significantly （P<0.01）. In terms of the relationship between ferroptosis and autophagy， compared with the blank group， the hormone group had significantly increased expression levels of LC3B， sTfR， Beclin1， and FTL and significantly decreased expression levels of GPX4 （P<0.01）. Compared with the hormone group， The ICA group had significantly decreased expression levels of LC3B， sTfR， and FTL and significantly increased expression levels of Beclin 1 and GPX4 （P<0.01）. Compared with those in the ICA group， the expression levels of LC3B， sTfR， and FTL increased in the rapamycin group， and those of Beclin 1 and GPX4 decreased （P<0.01）. In terms of cell ferroptosis and apoptosis，compared with the blank group， the hormone group had significantly increased expression levels of FTL， sTfR and Caspase-3 and significantly decreased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with the hormone group， the ICA group had significantly decreased expression levels of FTL， sTfR and Caspase-3 and significantly increased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with those in the ICA group， the expression levels of FTL， sTfR and Caspase-3 in the ferroptosis agonist group were increased， and the expression levels of GPX4， and Bcl-2 were decreased （P<0.01）. In terms of cell function，compared with that in the blank group， the ability of cell migration and tube formation was significantly decreased in the hormone group （P<0.01）. Compared with that in the hormone group， the cell migration and tube formation ability in the ICA group were significantly increased （P<0.01）. ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs， and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.

Objective:To investigate the mosquito-borne viruses carried by mosquito specimens collected from Huachuan county and Huanan county in Heilongjiang province.Methods:Mosquito samples were collected locally in 2023 and processed in the laboratory. Homogenates of the mosquitoes were inoculated into cells for virus isolation, followed by molecular and bioinformatics analyses of the viral isolates.Results:In 2023, ten viral isolates were obtained from Anopheles sinensis specimens collected in Heilongjiang province, China. Among these isolates, one was identified as Culex flavivirus (CxFV), one as Menghai rhabdovirus (MRV), and eight as Nam Dinh virus (NDiV). The phylogenetic analysis showed that CxFV belongs to genotype I and is clustered with the strains isolated from Liaoning province in 2011 and Ningxia Hui autonomous Region in 2019 in the same evolutionary branch, with amino acid similarity ranging from 98.2% to 99.2% and nucleotide similarity ranging from 98.8% to 99.2%. The MRV strain belongs to the same evolutionary subclade as the strain detected in Guangdong, with both nucleotide and amino acid similarity of 98.0%. Eight NDiV isolates clustered with the South Korean isolates on the same evolutionary branch, forming an independent evolutionary sub-branch. The nucleotide similarity among these eight isolates ranged from 98.5% to 99.7%, while the amino acid similarity ranged from 98.1% to 99.7%. In comparison, when matched with other NDiV isolates from China, the nucleotide similarity of these eight isolates ranged from 94.1% to 97.8%, and the amino acid similarity ranged from 93.5% to 97.7%.Conclusions:This study represents the first isolation of CxFV, MRV, and NDiV in Heilongjiang province, China, and the findings provide fundamental data for the prevention and control of mosquito-borne viral diseases in this region.

Objective:To establish a method for the rapid detection of varicella-zoster virus (VZV) by recombinase-aid amplification (RAA) combined with Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system.Methods:Clinical samples of suspected herpes zoster in Shandong province and Shanghai from 2023 to 2024 were collected, nucleic acids of positive samples were extracted, RAA-specific primers and crRNA (CRISPR RNA, crRNA) were designed for the conserved region of VZV, and the fluorescence intensity generated by Cas12a non-specific cleavage of single-stranded fluorescent probes was used to screen highly sensitive crRNAs and optimize the concentrations of crRNA, Cas12a and ssDNA probes. The sensitivity and reproducibility of the RAA-CRISPR/Cas12a detection method were evaluated by using synthesized plasmids and clinical samples, and the specificity of the method was evaluated by using other viral nucleic acids. The method was used to detect clinical samples by using the method and quantitative real-time PCR (qPCR) method, and the detection rate and consistency of the two method were compared.Results:The highly sensitive crRNA-4 was screened from the four crRNAs designed, and a VZV detection method for RAA-CRISPR/CAS12a based on fluorescence intensity measurement was established, which could be detected at 37℃ in 45 min, and the sensitivity of the detection could reach 10 copies/μL, a minimum clinical sample with a Ct value of 38.980 can be detected. It has high specificity and no cross-reactivity with Adenovirus 7, Herpes simplex virus type I, Herpes simplex virus type II, Coxsackieviruses A16, Cytomegalovirus, Epstein-Barr virus, Measles virus, Mumps virus, Enterovirus 71, Japanese encephalitis virus genotype 5. It has good stability, and can be successfully detected in low, medium and high concentrations of viral positive plasmids with good consistency. The detection rate of the clinically positive samples was 100%, which was completely consistent with the qPCR test result.Conclusions:RAA isothermal amplification technology combined with CRISPR-CAS12a technology was used to establish an accurate method for the detection of VZV virus, which was highly sensitive, specific, and had low requirements for experimental conditions, and could be completed within 45 min, which could provide strong technical support for the early detection of VZV.

Objective:To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the fast determination of maduramicin ammonium (MAD) in rat serum.Methods:In February 2024, rat serum samples were selected and directly injected after extraction and purification with methanol: acetonitrile (1: 1), separated on a C18 chromatographic column, and gradient-eluted using 0.1% formic acid aqueous solution -0.1% formic acid methanol solution as the mobile phase. Under optimized instrument conditions, electrospray positive ion multiple reaction monitoring (MRM) mode was employed for quantification using the external standard method, followed by methodological validation of the established approach.Results:The linearity of MAD in serum was good in the concentration range of 0.5-100 μg/L, and the correlation coefficient was 0.9997. The mean recoveries of MAD from spiked samples were 86.0%-109.6%, with the relative standard deviations were less than 10%. The limit of detection was 0.23 μg/L, The limit of detection was 0.75 μg/L.Conclusion:This method is high sensitive and reliable, which is suitable for the determination of MAD in in mouse serum.

Objective:To evaluate the effect of robot-assisted laparoscopic technology via abdominal approach for patients with primary retroperitoneal tumors.Methods:A retrospective cohort analysis was conducted for the clinical data of 71 patients who underwent robot-assisted laparoscopic resection of primary retroperitoneal tumor via abdominal approach at the Department of Urology of Sir Run Run Shaw Hospital,Zhejiang University School of Medicine from January 2015 to December 2023. There were 35 male and 36 female patients. The age ( M(IQR)) was 56(21) years (range: 21 to 83 years). The median tumor diameter was 46 (31) mm (range: 15 to 134 mm). Postoperative pathology revealed 58 benign and 13 malignant cases. Patients were divided into non-adherent group ( n=47) and adherent group ( n=24) based on whether the tumor was adhered to major organs or vessels. Perioperative and postoperative situation were compared between the two groups. Data comparisons were conducted using independent samples t-test for normally distributed continuous variables, Mann-Whitney U tests for non-normally distributed data, χ2 test or Fisher′s exact test for categorical variables. Kaplan-Meier survival analysis was employed to estimate 3-year recurrence or metastasis rate and 3-year mortality rate. Results:Operative time was 120(60) minutes (range: 45 to 440 minutes), intraoperative blood loss was 50 (80) ml (range: 10 to 2 000 ml). The median change of intraoperative mean arterial pressure was 40 (19) mmHg(1 mmHg=0.133 kPa)(range: 10 to 112 mmHg). Intraoperative blood transfusion was required in 7 cases, whereas 64 cases did not necessitate transfusion. The change in hemoglobin levels before and after surgery was (17.9±13.6) g/L (range:-16 to 53 g/L), and the median change in serum creatinine levels was 2.0 (14.5) μmol/L (range:-71.0 to 100.4 μmol/L). Postoperative fasting duration was 2.0 (1.5) days (range: 1 to 6 days), and the median hospital stay was 10.0 (7.5) days (range: 4 to 24 days). No perioperative mortality occurred in any of the patients. The non-adherent group had shorter operation time, less estimated blood loss, lower blood transfusion rate, smaller delta value of hemoglobin before and after surgery, larger delta value of creatinine before and after surgery, fewer postoperative complications, shorter postoperative fasting time, and shorter length of hospital stay than the adherent group(all P<0.05), while there was no significant difference in mean arterial pressure fluctuation between the two groups ( P>0.05). Follow-up data were available for 69 patients, with a median follow-up duration of 39 (43) months (range: 4 to 88 months). Among these patients, 40 completed the 3-year follow-up. The 3-year recurrence or metastasis rate was 10.14%, and the 3-year mortality rate was 2.90%. Conclusions:Robot-assisted laparoscopic technology via abdominal approach for resection of primary retroperitoneal tumors is safe and feasible. It can also achieve secure surgical outcome for primary retroperitoneal tumors adherent to surrounding organs or vessels, albeit with increased surgical complexity and slower postoperative recovery compared to non-adherent cases.

Objective:To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the fast determination of maduramicin ammonium (MAD) in rat serum.Methods:In February 2024, rat serum samples were selected and directly injected after extraction and purification with methanol: acetonitrile (1: 1), separated on a C18 chromatographic column, and gradient-eluted using 0.1% formic acid aqueous solution -0.1% formic acid methanol solution as the mobile phase. Under optimized instrument conditions, electrospray positive ion multiple reaction monitoring (MRM) mode was employed for quantification using the external standard method, followed by methodological validation of the established approach.Results:The linearity of MAD in serum was good in the concentration range of 0.5-100 μg/L, and the correlation coefficient was 0.9997. The mean recoveries of MAD from spiked samples were 86.0%-109.6%, with the relative standard deviations were less than 10%. The limit of detection was 0.23 μg/L, The limit of detection was 0.75 μg/L.Conclusion:This method is high sensitive and reliable, which is suitable for the determination of MAD in in mouse serum.

Objective:To investigate the effect of hydrogen on the expressions of stromal interaction molecule 1 (STIM1) and Ca 2+-release-activated-Ca 2+ channel protein 1 (Orai1) and the protective effect of hydrogen on septic mice acute lung injury (ALI). Methods:Forty-eight male ICR mice were divided into sham operation group (Sham group), hydrogen control group (Sham+H 2 group), sepsis group (SS group) and hydrogen intervention group (SS+H 2 group) according to a random number table method, with 12 mice in each group. Sepsis mice model were established by cecal ligation and puncture (CLP). Sham group and Sham+H 2 group did not undergo CLP, other operations were the same as follow. Sham+H 2 group and SS+H 2 group received 1 hour inhalation of 2% H 2 at 1 hour and 6 hours after CLP or sham operation. At 24 hours after CLP, 6 mice in each group were sacrificed for observing pulmonary microvascular permeability after injecting Evans blue (EB) through tail vein. Other 6 mice in each group were sacrificed for obtaining fresh lung tissue to observe the lung pathological change and lung wet/dry (W/D) weight ratio. The protein expressions and distribution of STIM1 and Orai1 in lung tissue were detected by Western blotting and immunofluorescence staining. The mRNA expressions of STIM1 and Orai1 in lung tissue were detected by reverse transcriotion-polymerase chain reaction (RT-PCR). Coimmunoprecipitation was used to observe STIM1-Orai1 interaction. Finally, pulmonary microvascular endothelial cells (PMVEC) of mice were cultured in vitro and randomly divided into four groups for inoculation onto culture plates: Control group, rich hydrogen solution group (Control+H 2 group), lipopolysaccharide (LPS) group and rich hydrogen solution intervention group (LPS+H 2 group). PMVECs in Control group and LPS group were cultured in normal medium. Control+H 2 group and LPS+H 2 group were cultured in saturated hydrogen medium. LPS group and LPS+H 2 group were added with LPS at 5 μg/mL. Intracellular Ca 2+ ([Ca 2+]i) concentration of PMVECs were detected by Fluo-4/AM green dye. Results:Compared with Sham group, the pathological score and lung W/D ratio were significantly increased in SS group at 24 hours after CLP (pathological score: 11.00±1.41 vs. 1.00±0.63, lung W/D ratio: 7.63±0.52 vs. 3.45±0.58, both P < 0.05), the content of EB in the lung tissue was increased (μg/g: 0.16±0.02 vs. 0.09±0.02, P < 0.05). Compared with SS group, the pathological score and lung W/D ratio were decreased in SS+H 2 group (pathological score: 3.50±1.05 vs. 11.00±1.41, lung W/D ratio: 4.45±0.45 vs. 7.63±0.52, both P < 0.05), the content of EB in the lung tissue was decreased (μg/g: 0.13±0.02 vs. 0.16±0.02, P < 0.05), which prove that hydrogen can improve ALI caused by sepsis. Compared with Sham group, the protein and mRNA expressions of STIM1 and Orai1 were up-regulated in SS group (relative expression level of STIM1 protein: 3.08±0.32 vs. 1.00±0.00, relative expression level of STIM1 mRNA: 3.65±0.24 vs. 1.00±0.00, relative expression level of Orai1 protein: 3.63±0.23 vs. 1.00±0.00, relative expression level of Orai1 mRNA: 3.80±0.22 vs. 1.00±0.00, all P < 0.05), while the protein and mRNA expressions of STIM1 and Orai1 were down-regulated in SS+H 2 group compared with SS group (relative expression level of STIM1 protein: 1.78±0.13 vs. 3.08±0.32, relative expression level of STIM1 mRNA: 1.76±0.28 vs. 3.65±0.24, relative expression level of Orai1 protein: 1.92±0.22 vs. 3.63±0.23, relative expression level of Orai1 mRNA: 1.85±0.18 vs. 3.80±0.22, all P < 0.05). Coimmunoprecipitation staining results showed that there was no statistically significance in the association between STIM1 and Orai1 in Sham group and Sham+H 2 group. Compared with Sham group, the STIM1-Orai1 interaction was increased in SS group (relative expression level: 3.71±0.37 vs. 1.00±0.00, P < 0.05), while the STIM1-Orai1 interaction was decreased in SS+H 2 group compared with SS group (relative expression level: 2.17±0.29 vs. 3.71±0.37, P < 0.05). There were no statistically significant differences in various indicators between Sham group and Sham+H 2 group. In vitro, the intracellular [Ca 2+]i concentration in PMVECs was increased in LPS group compared with Control group using Fluo-4/AM green dye. The intracellular [Ca 2+]i concentration in PMVECs was decreased in LPS+H 2 group compared with LPS group. Conclusion:The protective effect of hydrogen on lung tissues in septic mice is related to the inhibition of STIM1, Orai1 and the interaction between them.

Objective:To investigate the effect of hydrogen on the expressions of stromal interaction molecule 1 (STIM1) and Ca 2+-release-activated-Ca 2+ channel protein 1 (Orai1) and the protective effect of hydrogen on septic mice acute lung injury (ALI). Methods:Forty-eight male ICR mice were divided into sham operation group (Sham group), hydrogen control group (Sham+H 2 group), sepsis group (SS group) and hydrogen intervention group (SS+H 2 group) according to a random number table method, with 12 mice in each group. Sepsis mice model were established by cecal ligation and puncture (CLP). Sham group and Sham+H 2 group did not undergo CLP, other operations were the same as follow. Sham+H 2 group and SS+H 2 group received 1 hour inhalation of 2% H 2 at 1 hour and 6 hours after CLP or sham operation. At 24 hours after CLP, 6 mice in each group were sacrificed for observing pulmonary microvascular permeability after injecting Evans blue (EB) through tail vein. Other 6 mice in each group were sacrificed for obtaining fresh lung tissue to observe the lung pathological change and lung wet/dry (W/D) weight ratio. The protein expressions and distribution of STIM1 and Orai1 in lung tissue were detected by Western blotting and immunofluorescence staining. The mRNA expressions of STIM1 and Orai1 in lung tissue were detected by reverse transcriotion-polymerase chain reaction (RT-PCR). Coimmunoprecipitation was used to observe STIM1-Orai1 interaction. Finally, pulmonary microvascular endothelial cells (PMVEC) of mice were cultured in vitro and randomly divided into four groups for inoculation onto culture plates: Control group, rich hydrogen solution group (Control+H 2 group), lipopolysaccharide (LPS) group and rich hydrogen solution intervention group (LPS+H 2 group). PMVECs in Control group and LPS group were cultured in normal medium. Control+H 2 group and LPS+H 2 group were cultured in saturated hydrogen medium. LPS group and LPS+H 2 group were added with LPS at 5 μg/mL. Intracellular Ca 2+ ([Ca 2+]i) concentration of PMVECs were detected by Fluo-4/AM green dye. Results:Compared with Sham group, the pathological score and lung W/D ratio were significantly increased in SS group at 24 hours after CLP (pathological score: 11.00±1.41 vs. 1.00±0.63, lung W/D ratio: 7.63±0.52 vs. 3.45±0.58, both P < 0.05), the content of EB in the lung tissue was increased (μg/g: 0.16±0.02 vs. 0.09±0.02, P < 0.05). Compared with SS group, the pathological score and lung W/D ratio were decreased in SS+H 2 group (pathological score: 3.50±1.05 vs. 11.00±1.41, lung W/D ratio: 4.45±0.45 vs. 7.63±0.52, both P < 0.05), the content of EB in the lung tissue was decreased (μg/g: 0.13±0.02 vs. 0.16±0.02, P < 0.05), which prove that hydrogen can improve ALI caused by sepsis. Compared with Sham group, the protein and mRNA expressions of STIM1 and Orai1 were up-regulated in SS group (relative expression level of STIM1 protein: 3.08±0.32 vs. 1.00±0.00, relative expression level of STIM1 mRNA: 3.65±0.24 vs. 1.00±0.00, relative expression level of Orai1 protein: 3.63±0.23 vs. 1.00±0.00, relative expression level of Orai1 mRNA: 3.80±0.22 vs. 1.00±0.00, all P < 0.05), while the protein and mRNA expressions of STIM1 and Orai1 were down-regulated in SS+H 2 group compared with SS group (relative expression level of STIM1 protein: 1.78±0.13 vs. 3.08±0.32, relative expression level of STIM1 mRNA: 1.76±0.28 vs. 3.65±0.24, relative expression level of Orai1 protein: 1.92±0.22 vs. 3.63±0.23, relative expression level of Orai1 mRNA: 1.85±0.18 vs. 3.80±0.22, all P < 0.05). Coimmunoprecipitation staining results showed that there was no statistically significance in the association between STIM1 and Orai1 in Sham group and Sham+H 2 group. Compared with Sham group, the STIM1-Orai1 interaction was increased in SS group (relative expression level: 3.71±0.37 vs. 1.00±0.00, P < 0.05), while the STIM1-Orai1 interaction was decreased in SS+H 2 group compared with SS group (relative expression level: 2.17±0.29 vs. 3.71±0.37, P < 0.05). There were no statistically significant differences in various indicators between Sham group and Sham+H 2 group. In vitro, the intracellular [Ca 2+]i concentration in PMVECs was increased in LPS group compared with Control group using Fluo-4/AM green dye. The intracellular [Ca 2+]i concentration in PMVECs was decreased in LPS+H 2 group compared with LPS group. Conclusion:The protective effect of hydrogen on lung tissues in septic mice is related to the inhibition of STIM1, Orai1 and the interaction between them.

Objective:To investigate the mosquito-borne viruses carried by mosquito specimens collected from Huachuan county and Huanan county in Heilongjiang province.Methods:Mosquito samples were collected locally in 2023 and processed in the laboratory. Homogenates of the mosquitoes were inoculated into cells for virus isolation, followed by molecular and bioinformatics analyses of the viral isolates.Results:In 2023, ten viral isolates were obtained from Anopheles sinensis specimens collected in Heilongjiang province, China. Among these isolates, one was identified as Culex flavivirus (CxFV), one as Menghai rhabdovirus (MRV), and eight as Nam Dinh virus (NDiV). The phylogenetic analysis showed that CxFV belongs to genotype I and is clustered with the strains isolated from Liaoning province in 2011 and Ningxia Hui autonomous Region in 2019 in the same evolutionary branch, with amino acid similarity ranging from 98.2% to 99.2% and nucleotide similarity ranging from 98.8% to 99.2%. The MRV strain belongs to the same evolutionary subclade as the strain detected in Guangdong, with both nucleotide and amino acid similarity of 98.0%. Eight NDiV isolates clustered with the South Korean isolates on the same evolutionary branch, forming an independent evolutionary sub-branch. The nucleotide similarity among these eight isolates ranged from 98.5% to 99.7%, while the amino acid similarity ranged from 98.1% to 99.7%. In comparison, when matched with other NDiV isolates from China, the nucleotide similarity of these eight isolates ranged from 94.1% to 97.8%, and the amino acid similarity ranged from 93.5% to 97.7%.Conclusions:This study represents the first isolation of CxFV, MRV, and NDiV in Heilongjiang province, China, and the findings provide fundamental data for the prevention and control of mosquito-borne viral diseases in this region.

Objective:To establish a method for the rapid detection of varicella-zoster virus (VZV) by recombinase-aid amplification (RAA) combined with Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system.Methods:Clinical samples of suspected herpes zoster in Shandong province and Shanghai from 2023 to 2024 were collected, nucleic acids of positive samples were extracted, RAA-specific primers and crRNA (CRISPR RNA, crRNA) were designed for the conserved region of VZV, and the fluorescence intensity generated by Cas12a non-specific cleavage of single-stranded fluorescent probes was used to screen highly sensitive crRNAs and optimize the concentrations of crRNA, Cas12a and ssDNA probes. The sensitivity and reproducibility of the RAA-CRISPR/Cas12a detection method were evaluated by using synthesized plasmids and clinical samples, and the specificity of the method was evaluated by using other viral nucleic acids. The method was used to detect clinical samples by using the method and quantitative real-time PCR (qPCR) method, and the detection rate and consistency of the two method were compared.Results:The highly sensitive crRNA-4 was screened from the four crRNAs designed, and a VZV detection method for RAA-CRISPR/CAS12a based on fluorescence intensity measurement was established, which could be detected at 37℃ in 45 min, and the sensitivity of the detection could reach 10 copies/μL, a minimum clinical sample with a Ct value of 38.980 can be detected. It has high specificity and no cross-reactivity with Adenovirus 7, Herpes simplex virus type I, Herpes simplex virus type II, Coxsackieviruses A16, Cytomegalovirus, Epstein-Barr virus, Measles virus, Mumps virus, Enterovirus 71, Japanese encephalitis virus genotype 5. It has good stability, and can be successfully detected in low, medium and high concentrations of viral positive plasmids with good consistency. The detection rate of the clinically positive samples was 100%, which was completely consistent with the qPCR test result.Conclusions:RAA isothermal amplification technology combined with CRISPR-CAS12a technology was used to establish an accurate method for the detection of VZV virus, which was highly sensitive, specific, and had low requirements for experimental conditions, and could be completed within 45 min, which could provide strong technical support for the early detection of VZV.